Journal: Cell reports

Article Title: Principles of assembly and regulation of condensates of Polycomb repressive complex 1 through phase separation

doi: 10.1016/j.celrep.2023.113136

Figure Lengend Snippet: (A) A hypothetical model describing how condensate composition regulates the partitioning of CBX2-PRC1 components and nucleosomes and the exchange properties of the scaffold CBX2. Colored hexagons are the CBX2-PRC1 clients (magenta) and nucleosomes (green). (B) Representative epi-fluorescence images of CBX2-PRC1 subunits in the four-component (CBX2, RING1B [R], MEL18 [M], and PHC1 [P]) system. Scale bars, 5.0 μm. (C) Box plot of condensed fraction in the four-component system quantified from (B). p value is calculated using Student’s t test (*p < 0.05; **p < 0.01). (D) FRAP curves of CBX2 in the single-component, two-component, three-component, and four-component systems. Error bars denote SD. (E) Example confocal fluorescence images of CBX2 and nucleosomes (Nuc.) in the two-component, three-component, four-component, and five-component systems. Scale bars, 5.0 μm. (F) Box plot of condensed fraction of CBX2 and nucleosomes quantified from (E). p value is calculated using Student’s t test (**p < 0.01). (G) FRAP curves of YFP-CBX2 in the two-, three-, four-, and five-component systems. Error bars denote SD. (H) Representative live-cell epi-fluorescence images of HT-CBX2 in wild-type (WT), Ring1a −/− /b −/− , and Bmi1 −/− /Mel18 −/− mESC lines. Scale bars, 5.0 μm. (I and J) Box plots of condensed fraction (I) and size (J) of HT-CBX2 condensates quantified from (H). p value is calculated using Student’s t test (**p < 0.01). Error bars denote SD. (K) Example confocal images of FRAP of HT-CBX2 in wild-type (WT), Ring1a −/− /b −/− , and Bmi1 −/− /Mel18 −/− mESC lines. Red arrows show condensates to be bleached. Scale bar, 5.0 μm. (L) FRAP curves of HT-CBX2 within and outside condensates in wild-type (WT), Ring1a −/− /b −/− , and Bmi1 −/− /Mel18 −/− mESC lines. Error bars denote SD.

Article Snippet: The FRAP curves were fitted by a single-exponential function implemented in OriginLab as described in. (Equation 5) I = r − m ⋅ e ( − k ⋅ t ) Where m is the mobile fraction.

Techniques: Fluorescence

Journal:

Article Title: Regulation of CFTR Trafficking by Its R Domain * S⃞

doi: 10.1074/jbc.M800516200

Figure Lengend Snippet: The functional half-life of CFTR is reduced during cAMP/PKA stimulation. Current decay curves were produced by continuous incubation of oocytes expressing WT-CFTR (1 ng) and β2-adrenergic receptor (1 ng) with brefeldin A (10 μm). The data were fit with a single exponential function using SigmaPlot (Systat Software). A, oocytes were stimulated continuously with 10 μm isoproterenol, and currents were recorded at 0, 2, 4, 8, and 24 h (N = 5; n = 20, each data point). B, currents were recorded from oocytes at separate time points (0, 1, 2, 4, 8, and 24 h) after initial treatment with 10 μm BFA. Current measurements were obtained at maximal stimulation following isoproterenol stimulation at the indicated times by arrowheads (N = 4; n = 16).

Article Snippet: The data were fit with a single exponential function using SigmaPlot (Systat Software).

Techniques: Functional Assay, Produced, Incubation, Expressing, Software